1. CHARACTERISATION OF NEW PARTS

Our pTAC with RFP reporter was characterised at lab scale by carrying out a successful transformation with a control. By transforming the parts into competent cells and adding IPTG to the non-control plate, we expected to see the control's colour remain unchanged whilst that of the other plate to turn red indicating that the IPTG will have induced the activation of the pTAC and thus characterise our part. Obviously, we had hoped to be in a position to characterise the whole genetic circuit but due to time restraints we could not complete our circuit.

Check out our parts page to see the parts submitted in more detail UCL_London 2010 iGEM Team Parts.

IPTG induction of the reporter device BBa_K301001 was demonstrated in a lacI host strain with 0.1mM IPTG (panel B). A lac promoter device was tested as positive control (panel C).

As predicted, IPTG induction was not observed when our new device resided in a lacI negative host strain (panel A) or when the pTac biobrick was omitted (panel D).

2. CHARACTERISATION OF EXISTING PARTS/DEVICES AT 1 L FERMENTER SCALE

As part of the Gold criteria which is our target, we had to characterise an existing part, and so we decided to characterise 3 devices from our 2009 parts;

Fermenter 1 - UCL_London 2009 iGEM BBa_K239011 PART

GFP test devise for Spy promoter

http://partsregistry.org/wiki/index.php?title=Part:BBa_K239011

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Fermenter 2 -UCL_London 2009 iGEM BBa_K239009 PART

GFP test devise for DegP promoter

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http://partsregistry.org/wiki/index.php?title=Part:BBa_K239009

Fermenter 3 - UCL_London 2009 iGEM BBa_K239015 PART

Stress Light GFP Anaerobic Metabolism Detector

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The 3 parts were inoculated into separate 1 L fermenters and we ran the processes in parallel along side a control process. We observed the various stages of the process from the growth to the lag phase, and what we hoped to observe was the progressive change of the colours of the 3 processes into green. As we expected, the induction of the GFP protein on addition of IPTG caused the induction of the promoter and thus the green proteins release into the culture.

Control Variable Test Variable

The following video shows the moment of that our iGEM aspirations came true, and we saw green fluorescence;

From the paste removed from the fermenter vessel following the successful characterisation of the parts, we used it to creatively mark the words iGEM in plates and hence further emphasizing the results that the parts do work.